scholarly journals Detection of Zoonotic Yersinia Infection in Symptomatic and Asymptomatic Dogs in Sokoto State Nigeria

2020 ◽  
Vol 1 (2) ◽  
Author(s):  
Nicholas Nathaniel Pilau ◽  
Shehu Zaid ◽  
Abubakar Sadiq Yakubu ◽  
Bashir Saidu ◽  
Umar Yakubu Dabai
Keyword(s):  
Selective Medium ◽  
Biochemical Tests ◽  
Selective Media ◽  
Yersinia Species ◽  
Yersinia Infection ◽  
Animal Populations ◽  
Pure Cultures ◽  
Animal Hosts ◽  
Relative Prevalence ◽  
Suspicion Index

Background: Zoonotic Yersinia infection has been previously reported in humans and animal hosts in Nigeria, occasionally with fulminant disease. Despite earlier evidence of Yersinia pathogen circulating in human and animal populations in Nigeria, studies and suspicion index to Yersinia is below an acceptable average amongst clinicians, diagnosticians, academics and health policy officers. Methods: The Deoxycholate Citrate Agar (DCA) was used as selective media to culture Yersinia preceded by inoculation in MacConkay agar. Plates with evident growth in the differential media consistent with reported accounts for Yersinia were picked and inoculated in selective medium and left for 48 hours until growth was seen, other samples left until five days before being discarded as negative. Pure cultures were subjected to comprehensive biochemical tests standard and previously applied for diagnosis and discrimination of Yersinia species. Result: This research recorded an overall microbial prevalence of 30%. Prevalence of Yersinia enterocolitica was 18.3% and Y. pseudotuberculosis 11.7%. Male dogs presented a relative prevalence of Y.

Detection of Zoonotic Yersinia Infection in Symptomatic and Asymptomatic Dogs in Sokoto State Nigeria

2020 ◽  
Vol 1 (2) ◽  
Author(s):  
Nicholas Nathaniel Pilau ◽  
Shehu Zaid ◽  
Abubakar Sadiq Yakubu ◽  
Bashir Saidu ◽  
Umar , Yakubu Dabai ,
Keyword(s):  
Health Policy ◽  
Selective Medium ◽  
Selective Media ◽  
Yersinia Species ◽  
Yersinia Infection ◽  
Animal Populations ◽  
Pure Cultures ◽  
Animal Hosts ◽  
Relative Prevalence ◽  
Suspicion Index

Background: Zoonotic Yersinia infection has been previously reported in humans and animal hosts in Nigeria, occasionally with fulminant disease. Despite earlier evidence of Yersinia pathogen circulating in human and animal populations in Nigeria, studies and suspicion index to Yersinia is below an acceptable average amongst clinicians, diagnosticians, academics and health policy officers. Methods: The deoxycholate Citrate Agar (DCA) was used as a selective media to culture Yersinia, preceded by inoculation in MacConkay agar. Plates with evident growth in the differential media consistent with reported accounts for Yersinia were picked and inoculated in selective medium and left for 48 hours until growth was seen, other samples were left until five days before being discarded as negative. Pure cultures were subjected to a comprehensive biochemical test standard and previously applied for diagnosis and discrimination of Yersinia species. Result: This research recorded an overall microbial prevalence of 30%. Prevalence of Yersinia enterocolitica was 18.3% and Y. pseudotuberculosis 11.7%. Male dogs presented a relative prevalence of Y. enterocolitica 40.9% compared with 59.1% recorded for female dogs. Symptomatic dogs presented a relative prevalence of Y. enterocolitica of 86.4% and Y. pseudotuberculosis of 71.4%

scholarly journals MOLECULAR DETECTION AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM RAW MILK SOLD IN DIFFERENT MARKETS OF BANGLADESH

2017 ◽  
Vol 14 (2) ◽  
pp. 277-282 ◽  
Author(s):  
M. A. Islam ◽  
S. M. L. Kabir ◽  
M. T. Rahman
Keyword(s):  
Molecular Detection ◽  
Culture Media ◽  
Raw Milk ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Selective Media ◽  
Milk Samples ◽  
Antimicrobial Sensitivity ◽  
Antimicrobial Sensitivity Test

The study was intended for molecular detection of S. aureus isolated from raw cow’s milk. A total of 20 milk samples were collected from different upazila markets of Jamalpur, Tangail, Kishoreganj and Netrokona districts of Bangladesh. Milk samples were cultured onto various culture media for the isolation of bacteria. The isolated bacteria were identified by studying cultural properties on different selective media, biochemical tests, and finally by PCR. Out of 20 samples, 15 (75%) milk samples were found to be positive for S. aureus. S. aureus specific 16S rRNA gene was amplified from all isolates and identified as S. aureus. Antimicrobial sensitivity test was carried out to ascertain the susceptibility of the organism to various antibiotics. Its results showed that the S. aureus isolates were resistant to amoxicillin (100%), erythromycin (73.33%) and tetracycline (73.33%) but sensitive to azithromycin (93.33%), ciprofloxacin (93.33%), gentamicin (100%), norfloxacin (86.67%) and streptomycin (86.67%).

Use of Oligonucleotide Array for Identification of Six Foodborne Pathogens and Pseudomonas aeruginosa Grown on Selective Media

2005 ◽  
Vol 68 (11) ◽  
pp. 2278-2286 ◽  
Author(s):  
MIAO CHU LIN ◽  
AY HUEY HUANG ◽  
HAU YANG TSEN ◽  
HIN-CHUNG WONG ◽  
TSUNG CHAIN CHANG
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Foodborne Pathogens ◽  
Intergenic Spacer ◽  
Oligonucleotide Array ◽  
Universal Primers ◽  
Morphological Observation ◽  
Biochemical Tests ◽  
Nylon Membrane ◽  
Selective Media ◽  
Species Specific

Identification of presumptive foodborne pathogens grown on selective media may take one to several days and requires a different battery of biochemical tests for each microorganism. A molecular identification method was developed in which universal primers were used to amplify the 16S to 23S rDNA intergenic spacer of target microorganisms, and PCR products were hybridized to a panel of species-specific oligonucleotides that were immobilized on a nylon membrane. The seven target microorganisms were Bacillus cereus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella, Staphylococcus aureus, and Vibrio parahaemolyticus. After testing a large collection of target bacteria (29 to 51 strains) and nontarget bacteria (>500 strains), the performances (sensitivity and specificity) of the oligonucleotide array were as follows: B. cereus (100 and 77%), E. coli (100 and 100%), L. monocytogenes (100 and 90%), P. aeruginosa (100 and 100%), Salmonella (100 and 100%), S. aureus (100 and 100%), and V. parahaemolyticus (100 and 94.2%). Other species in the B. cereus group cross-hybridized to the probes used for identification of B. cereus, and positive results should be confirmed by additional morphological observation of colonies. Listeria innocua cross-reacted with probes used to identify L. monocytogenes, but a simple hemolysis test was used to differentiate the two species. Some strains of Vibrio harveyi and Vibrio mimicus cross-hybridized with probes used for identification of V. parahaemolyticus and caused false-positive reactions. The advantage of the array is that a common protocol was used to identify the seven target microorganisms and multiple different microorganisms could be simultaneously identified on a single array.

Enumeration and Identification of Coliform Bacteria Injured by Chlorine or Fungicide Mixed with Agricultural Water

2016 ◽  
Vol 79 (10) ◽  
pp. 1789-1793 ◽  
Author(s):  
HIDEMI IZUMI ◽  
YUJI NAKATA ◽  
AYANO INOUE
Keyword(s):  
Cell Injury ◽  
Pond Water ◽  
Coliform Bacteria ◽  
Agricultural Water ◽  
Isolation And Identification ◽  
Electrolyzed Water ◽  
Selective Media ◽  
E Coli ◽  
Pure Cultures ◽  
Water Pond

ABSTRACT Chemical sanitizers may induce no injury (bacteria survive), sublethal injury (bacteria are injured), or lethal injury (bacteria die). The proportion of coliform bacteria that were injured sublethally by chlorine and fungicide mixed with agricultural water (pond water), which was used to dilute the pesticide solution, was evaluated using the thin agar layer (TAL) method. In pure cultures of Enterobacter cloacae, Escherichia coli, and E. coli O157:H7 (representing a human pathogen), the percentage of chlorine-injured cells was 69 to 77% for dilute electrolyzed water containing an available chlorine level of 2 ppm. When agricultural water was mixed with electrolyzed water, the percentage of injured coliforms in agricultural water was 75%. The isolation and identification of bacteria on TAL and selective media suggested that the chlorine stress caused injury to Enterobacter kobei. Of the four fungicide products tested, diluted to their recommended concentrations, Topsin-M, Sumilex, and Oxirane caused injury to coliform bacteria in pure cultures and in agricultural water following their mixture with each pesticide, whereas Streptomycin did not induce any injury to the bacteria. The percentage of injury was 45 to 97% for Topsin-M, 80 to 87% for Sumilex, and 50 to 97% for Oxirane. A comparison of the coliforms isolated from the pesticide solutions and then grown on either TAL or selective media indicated the possibility of fungicide-injured Rahnella aquatilis, Yersinia mollaretii, and E. coli. These results suggest the importance of selecting a suitable sanitizer and the necessity of adjusting the sanitizer concentration to a level that will kill the coliforms rather than cause sanitizer-induced cell injury that can result in the recovery of the coliforms.

scholarly journals Comparison of Six Commercially Available Transport Media for Maintenance of Serpulina (Treponema) Hyodysenteriae

1992 ◽  
Vol 4 (3) ◽  
pp. 285-292 ◽  
Author(s):  
G. E. Duhamel ◽  
R. J. Bernard ◽  
M. R. Mathiesen ◽  
K. M. Eskridge
Keyword(s):  
Pure Culture ◽  
Selective Medium ◽  
Fecal Flora ◽  
Transport Medium ◽  
Selective Transport ◽  
Incubation Periods ◽  
Pure Cultures ◽  
Treponema Hyodysenteriae ◽  
Better Than ◽  
Anaerobic Media

Two anaerobic (A1 and A2), 1 selective (S1), and 3 conventional (C1, C2, and C3) transport media formulations were compared for their capacity to maintain the viability of Serpulina (Treponema) hyodysenteriae. Initial experiments compared the recovery of S. hyodysenteriae from pure cultures held in each transport medium for 0.5, 1, 2, 3, 5, and 7 days at −40 C, 4 C, 25 C, and 36 C. Subsequent experiments compared each transport medium for maintenance of S. hyodysenteriae in fecal specimens obtained from experimentally infected pigs after holding for up to 7 days at 25 C. In each experiment, the viability of S. hyodysenteriae in each transport medium incubated at each temperature and for each period was determined by inoculating the transport medium onto either trypticase soy agar with 5% sheep blood or selective BJ agar and incubating at 42 C anaerobically. Viability and fecal flora contamination were evaluated blindly after 2-, 4-, and 6-day incubation periods. At −40 C, recovery of viable S. hyodysenteriae from pure culture did not differ among the transport media from 0.5 to 7 days, and all of the transport media consistently maintained the viability of the spirochetes for 7 days. At 4 C, the anaerobic and selective transport media maintained the viability of pure cultures of S. hyodysenteriae significantly better than did the conventional transport media group at day 7 ( P = 0.019). At the same temperature, the anaerobic media maintained viability better than did the conventional media at 5 days ( P < 0.042). At 25 C, the anaerobic transport media were significantly better than the conventional transport media at maintaining the viability of pure cultures of the spirochetes at 2, 3, and 5 days ( P < 0.018) and were significantly better than the selective medium at 3 days (P = 0.012). At 36 C, the recovery of viable spirochetes was significantly better with the anaerobic transport media than with both the conventional media for days 2–7 ( P < 0.006) and the selective medium for days 3–7 ( P < 0.049). Fecal specimens held in transport media Al and C1, as a group, had significantly higher viability than those held in the other transport media formulations taken as a group at all incubation times, except 0.5 day ( P < 0.0046). Contamination of selective BJ medium by fecal flora was markedly higher after holding fecal specimens in conventional transport media than in anaerobic and selective transport media. In a dilution trial of a pure culture of spirochetes, transport media Al and A2 maintained the viability of 108 S. hyodysenteriae for 7 days; however, medium Al was 10–100-fold more effective than medium A2 when lower initial concentrations of spirochetes were sampled. In a dilution trial of a fecal specimen, medium Al maintained the viability of 101 spirochetes for 2.5 days compared with 103 with medium C1. Overall, media A1 and C1 were the most satisfactory transport media for recovery of viable S. hyodysenteriae from fecal specimens held at 25 C for up to 7 days.

scholarly journals Molecular characterization of covalently closed circular plasmid DNA in local Streptomyces strains isolated from soil with detection of antibiotic production and antibacterial activity

2019 ◽  
Vol 10 (3) ◽  
pp. 2142-2152
Author(s):  
Suad A Al-Hilu ◽  
Wisam H Al-Shujairi
Keyword(s):  
Antibacterial Activity ◽  
Yeast Extract ◽  
Plasmid Dna ◽  
Antibiotic Production ◽  
Selective Medium ◽  
Malt Extract ◽  
Biochemical Tests ◽  
Alkaline Lysis ◽  
Mannitol Salt Agar ◽  
Streptomyces Species

Various of Streptomyces species have two kinds of plasmids, circular plasmids (8 to 31 kb) and linear plasmids (12 to 1700 kb). Covalently closed circular (CCC) plasmids are profuse in the genus of Streptomyces and involved in production and resistance of antibiotics by genetic controlling. We collected fifty clinical soil samples from different regions in Al-Najaf Al-Ashraf province/Iraq. The samples included five from Al-Ghadeer Quarter, five from Al-Karama Quarter, 10 from Kufa University, five from Al-Ameer Quarter, four from Al-Forat Quarter, 10 from North Quarters and eleven from desert roads in Al-Najaf. Diluted samples were cultured on Yeast extract Malt extract (YEME) agar medium as a selective medium; then the presumptive Streptomyces colonies were subcultured on Tryptone Yeast extract (TYE) agar, then incubation at 37ᵒC for 7 days. Seven biochemical tests for identification of Streptomyces isolates these are: Catalase test, Oxidase test, Urase test, Kligler Iron Agar test (KIA), Simmon᾽s Citrate test, addition to MacConkey agar test and Mannitol Salt agar test. Five antibiotic discs were used for detection of antibiotic sensitivity of the Streptomyces isolates; these are: Tetracycline, Gentamycin, Vancomycin, Ampicillin, Erythromycin. The sensitivity of the antibiotics was observed by recorded the diameter of inhibition zone around the discs. Two test bacteria (Staphylococcus aureus and E. coli) were used for the determination of antibacterial activity. Plasmid isolation was done by the alkaline lysis method. This method is characterized by the rapid isolation of DNA from Streptomyces. Then, detection of Plasmid DNA occurred by using agarose gel electrophoresis.

scholarly journals First Report of Phytophthora palmivora on Coronilla valentina subsp. glauca in Italy

Plant Disease ◽  
2002 ◽  
Vol 86 (3) ◽  
pp. 327-327
Author(s):  
S. O. Cacciola ◽  
A. Pane ◽  
S. Davino ◽  
F. Raudino
Keyword(s):  
Mating Type ◽  
Irrigation System ◽  
Field Capacity ◽  
Industrial Area ◽  
Selective Medium ◽  
Phytophthora Palmivora ◽  
Dual Culture ◽  
Trickle Irrigation ◽  
First Report ◽  
Pure Cultures

The genus Coronilla L. (family Fabaceae), which includes several species native to central and southern Europe, such as C. varia L. (axseed or crown-vetch), C. emerus (scorpion senna), and C. valentina L., is used in Italy as a landscape shrub or potted ornamental plant. During the summer of 2001, 80% of approximately 10,000 1-year-old plants of C. valentina subsp. glauca (L.) Batt. used to landscape an industrial area in the Caltanissetta Province (Sicily) showed symptoms of dieback associated with basal stem and root rot. Plants had been transplanted from pots in April and watered using a trickle irrigation system. A species of Phytophthora was isolated consistently from rotted roots and basal stems using BNPRAH selective medium (3). Pure cultures of this fungus were obtained by single-hypha transfers. Ten isolates, each originating from a single plant, were identified as P. palmivora (Butler) Butler on the basis of morphological and cultural characters as described by Erwin and Ribeiro (1). On solid media, including potato dextrose agar, cornmeal agar, and V8-juice agar, all the isolates produced elliptical to ovoid, papillate sporangia with a mean length/breadth ratio of 1.8. Sporangia were caducous with a short pedicel (mean pedicel length = 5 µm) and a conspicuous basal plug. Mating type was determined on V8 agar in dual culture with mating type A1 and A2 of reference isolates of P. nicotianae and P. palmivora. All isolates were heterothallic and produced oogonia and oospores only with reference isolates of the A2 mating type. Antheridia were amphigynous. Electrophoresis of mycelial proteins on polyacrylamide slab gel confirmed that all isolates were pure cultures and belonged to the same species. Koch's postulates were fulfilled using 6-month-old C. valentina subsp. glauca plants that were transplanted into pots filled with soil artificially inoculated with chlamydospores (50 chlamydospores per gram of soil) produced in submerged axenic cultures (2). The plants were maintained in a glasshouse at temperatures ranging from 18 to 28°C, and the pots were watered to field capacity once a week. One month after transplanting, 70% of plants showed dieback symptoms, while control plants, which were grown in pots containing noninoculated soil, remained healthy. The pathogen was reisolated from roots and basal stems of symptomatic plants. These results demonstrate that P. palmivora is the causal agent of dieback of C. valentina subsp. glauca plants. High temperatures in summer and waterlogging of soil due to excess irrigation water could have enhanced disease development. To our knowledge, this is the first report of P. palmivora on a species of Coronilla. P. palmivora is an exotic pathogen, but it is becoming widespread in Italy, where it has been reported from various regions on different hosts, including cyclamen, English ivy, palms, Pittosporum, and olive. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St Paul, MN, 1996. (2) J. Y Kadooka and W. H. Ko. Phytopathology 63:559, 1973. (3) H. Masago et al. Phytopathology 67:425, 1977.

scholarly journals DETEKSI FENOTIPIK Methicillin Resistant Staphylococcus aureus (MRSA) PADA SAMPEL MAKANAN DI SIDOARJO

2019 ◽  
Vol 7 (1) ◽  
pp. 55-65
Author(s):  
Yulianto Ade Prasetya
Keyword(s):  
Clinical Laboratory ◽  
Bacterial Species ◽  
Antibiotic Sensitivity ◽  
Food Samples ◽  
Selective Medium ◽  
Beta Lactam ◽  
Biochemical Tests ◽  
Methicillin Resistant ◽  
Staphyloccocus Aureus ◽  
Fried Foods

Methicillin-resistant Staphyloccocus aureus (MRSA) is an isolate that is resistant to the antibiotic methicillin and beta lactam group. The incidence of MRSA associated with nosocomial infections in various parts of the world is very high, but research on its spread in community infections is rarely reported. This study aims to detect the presence of phenotypic MRSA in food samples in Sidoarjo. The food samples (cilok, fried foods and tempura) collected were then weighed, diluted, and cultured in a selective medium and differential namely Manitol Salt agar. The yellow-colored colonies were then continued with microscopic testing and biochemical tests to distinguish between Staphylococcus species. Thirty eight collected Staphyloccus aureus isolates were then screened using Oxacillin 1 µg and there were eight (8) isolates that were positive for MRSA according to the criteria of the Clinical Laboratory Standart Institutre (CLSI). Eight of the isolates were tested for antibiotic sensitivity with the Kirby-Bauer method with Chloramphenicol 30 µg and Cotrimoxazole 25 µg. Eight MRSA (21%) isolates were resistant to Chloramphenicol and only four isolates were resistant to Cotrimoxazole. The presence of MRSA isolates in community infections needs to be watched out for considering these genes can be transmitted and spread between bacterial species

scholarly journals Recovery and Enumeration of Staphylococcus aureus by the Selective Agar

Fine Focus ◽  
2016 ◽  
Vol 2 (1) ◽  
pp. 51-59
Author(s):  
Jill Bange ◽  
Emily Brumfield ◽  
Alysha L. Ellison
Keyword(s):  
Staphylococcus Aureus ◽  
Food Processing ◽  
Recovery Period ◽  
False Negative ◽  
Selective Medium ◽  
Bacterial Cells ◽  
Human Immune System ◽  
Selective Media ◽  
Selective Agar ◽  
Overlay Method

Staphylococcus aureus isan example of a commensal bacterium responsible for emesis, acute diarrheal syndrome, and sepsis. S. aureus often must be isolated from patient samples in a clinical setting or from food samples during food processing in an industrial setting, although these bacterial cells may be injured by the human immune system or by food processing measures. Therefore, injured cells may not be fully recovered on media selective for S. aureus and enumeration (e.g., CFU/mL) may not reflect the true concentration of the original sample. The objective of this study was to determine whether the selective agar overlay method of recovery is more sensitive, selective, and time-effective for enumeration of artificially injured S. aureus cultures when compared to more traditional techniques. The selective agar overlay method involves pour plating S. aureus in non-selective medium, allowing the sample to incubate for a four hour recovery period, and then overlaying selective medium over the non-selective medium. Artificial injury of S. aureus cells was accomplished by treatment with carvacrol, an extract from oil of oregano. Our results indicated that carvacrol-injured S. aureus cells were recovered by the selective agar overlay at the same concentration as recovery on non-selective media, and at a significantly higher concentration than recovery on selective media. This method allows for more rapid and accurate diagnoses, and may be more cost-effective due to the reduction or elimination of false negative results.

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